Brassinosteroids (BRs) as a class of steroid plant hormones participate in the regulation of numerous developmental processes, including root and shoot growth, vascular differentiation, fertility, flowering, and seed germination, as well as in responding to environmental stresses. During four decades of research, the BR biosynthetic pathways have been well studied with forward-and reverse genetics approaches. The free BRs contain 27, 28, and 29 carbons within their skeletal structure: (1): 5a-cholestane or 26nor-24a-methyl-5a-cholestane for C 27-BRs; (2) 24a-methyl-5a-cholestane, 24b-methyl-5a-cholestane or 24-methylene-5a-cholestane for C 28-BRs; (3) 24a-ethyl-5acholestane, 24(Z)-ethylidene-5a-cholestane, 25-methyl-5a-campestane or 24methylene-25-methyl-5a-cholestane for C 29-BRs, as well as different kinds and orientations of oxygenated functions in A-and Bring. These alkyl substituents are also common structural features of sterols. BRs are derived from sterols carrying the same side chain. The C 27-BRs without substituent at C-24 are biosynthesized from cholesterol. The C 28-BRs carrying either an a-methyl, b-methyl, or methylene group are derived from campesterol, 24-epicampesterol or 24-methylenecholesterol, respectively. The C 29-BRs with an a-ethyl group are produced from sitosterol. Furthermore, the C 29 BRs carrying methylene at C-24 and an additional methyl group at C-25 are derived from 24methylene-25-methylcholesterol. Generally, BRs are biosynthesized via cycloartenol and cycloartanol dependent pathways. Till now, more than 17 compounds were characterized as inhibitors of the BR biosynthesis. For nine of the inhibitors (e.g., brassinazole and YCZ-18) a specific target reaction within the BR biosynthetic pathway has been identified. Therefore, the review highlights comprehensively recent advances in our understanding of the BR biosynthesis, sterol precursors, and dependencies between the C 27-C 28 and C 28-C 29 pathways.